Abstract
Monoclonal antibodies (Mabs) are valuable reagents for the purification, characterization and immunolocalization of proteins.In this study, we raised Mabs against human peroxisome proliferator-activated receptors (PPARs) using baculovirus particles displaying surface glycoprotein gp64-fusion proteins as the immunizing agent.In this system, to display fusion proteins on the viral surface, the amino terminal sequences of human PPARd and PPARg2 are inserted in-frame between the signal sequence and the mature domain of the gp64 nucleotide sequence.Mabs were raised by immunization with whole virus without a purification of the target antigens.The Mabs generated by this novel method were shown to recognize not only the gp64-PPARs fusion protein, but also mature, expressed proteins by a wide variety of techniques, including immunohistochemistry, immunoblotting, and electrophoretic mobility shift assays (EMSAs).Transfection of the transfer vector containing a nucleotide sequence encoding less than 30 amino acids along with linearized baculovirus DNA allows for the production of a high affinity antibody against the corresponding mature form.This method is of potential utility in that it allows the production of valuable antibodies without the requirement of a protein purification step.
Original language | English (US) |
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Pages (from-to) | 233-242 |
Number of pages | 10 |
Journal | Journal of atherosclerosis and thrombosis |
Volume | 9 |
Issue number | 5 |
DOIs | |
State | Published - 2002 |
ASJC Scopus subject areas
- Internal Medicine
- Cardiology and Cardiovascular Medicine
- Biochemistry, medical